A. Effects of AEA on intracellular cAMP levels. BeWo cells were treated (± 10 μM AEA; ± 50 μM
forskolin, FOR) for 1 h and collected in 0.1 M HCI. Intracellular concentrations of cAMP were determined by ELISA and normalized
to protein concentration. Data are presented as mean ± SE (n = 4). B. 8-Br-cAMP abrogates AEA effects on BCRP mRNA. qPCR
was performed on BeWo cells treated with AEA (10 μM), 8-Br-cAMP (1 mM), or both for 24 h. Black and white bars represented
samples treated in the absence or presence of AEA, respectively. Data are presented as mean ± SE (n = 4). D. Inhibition of
BCRP efflux activity by AEA is cAMP-dependent. BeWo cells were treated with either AEA (10 μM), 8-Br-cAMP (1 mM), or both
for 24 h. Cells were then trypsinized and subjected to the Hoechst 33342 transport assay as described in Materials and Methods.
Data represent the mean ± SE (n = 3-5). Asterisks (*) represent statistically significant differences (p < 0.05)
compared to control cells. Daggers (†) represent statistically significant differences (p < 0.05) compared to
AEA-treated cells.